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摸索体外诱导真菌耐药突变菌落的经济、简便的方法，为进行耐药性研究提供实验模型。方法：对所收集的３４株新型隐球菌临床分离株参照美国国家临床实验室标准化委员会（ＮＣＣＬＳ）推荐的Ｍ２７－Ａ方案，采用微量稀释法测定了其氟康唑最低抑菌浓度（ＭＩＣ），选出２株敏感株（ＭＩＣ＝４ｕｇ／ｍｌ）ＨＳ１７０６１、ＨＳ１６５３２和１株标准株ＢＬＳ１０９作为实验原始菌株，经在不同浓度氟康唑的培养基中连续传代培养处理，诱导产生耐药突变菌落。结果：经传代培养，获得了抗不同浓度氟康唑的耐药突变菌落，且突变菌落氟康唑的耐药性是稳定的。结论：通过采用以不同氟康唑浓度梯度连续传代培养的方法可诱导产生氟康唑耐药株，遗传稳定性测定表明耐药突变菌落的抗氟康唑特性是稳定的，本方法可为进行耐药前后的对比研究提供可靠的实验模型。
To explore the economical and convenient method of inducing fungal drug-resistant mutant colonies in vitro, and to provide an experimental model for the study of drug resistance. Methods: Thirty-four clinical isolates of Cryptococcus neoformans were collected according to the M27-A protocol recommended by the National Committee for Clinical Laboratory Standards (NCCLS). The minimum inhibitory concentration (MIC) of fluconazole, Two sensitive strains (MIC = 4ug / ml), HS17061 and HS16532, and one strain of standard strain BLS109 were selected as the original strains. After continuous subculture in different concentrations of fluconazole, drug-resistant mutant colonies were induced. Results: After subculture, the resistant colonies resistant to fluconazole of different concentrations were obtained, and the susceptibility of the mutant colonies to fluconazole was stable. CONCLUSIONS: Fluconazole-resistant strains can be induced by continuous subculturing with different concentrations of fluconazole and the genetic stability assay shows that the anti-fluconazole characteristics of the drug-resistant mutant colonies are stable and the method can be performed Comparative studies before and after resistance provide a reliable experimental model.